Halocatena marina sp. nov., a novel filamentous halophilic archaeon isolated from marine tidal flat and emended description of the genus Halocatena.

Three novel filamentous halophilic archaea, strains DFN5T, RDMS1, and QDMS1, were isolated from the coastal saline soil samples of the intertidal zones located in different regions of Jiangsu Province, China. The colonies of these strains were pinkish-white due to the presence of white spores. These three strains are extremely halophilic and grew optimally at 35-37 °C and pH 7.0-7.5. Based on 16S rRNA and rpoB' gene analysis, strains DFN5T, RDMS1, and QDMS1 gathered together in phylogenetic trees and then clustered with the current species of the genus Halocatena showing 96.9-97.4% and 82.2-82.5% similarities, respectively. Both the 16S rRNA gene-based and rpoB' gene-based phylogenies were fully supported by the phylogenomic analysis, and the overall genome-related indexes indicated that strains DFN5T, RDMS1, and QDMS1 should be a novel species of the genus Halocatena. Genome mining revealed that there are considerable differences in the genes related to ß-carotene synthesis among these three strains and the current species of Halocatena. The major polar lipids of strains DFN5T, RDMS1, and QDMS1 are PA, PG, PGP-Me, S-TGD-1, TGD-1, and TGD-2. The minor polar lipids, S-DGD-1, DGD-1, S2-DGD, and S-TeGD may be detected. According to the phenotypic characteristics, phylogenetic analysis, genomic and chemotaxonomic features, strains DFN5T (= CGMCC 1.19401 T = JCM 35422 T), RDMS1 (= CGMCC 1.19411) and QDMS1 (= CGMCC 1.19410) were classified as a novel species of the genus Halocatena with the proposed name, Halocatena marina sp. nov. This is the first report of the description of a novel filamentous haloarchaeon isolated from marine intertidal zones.

Hly176B, a low-salt tolerant halolysin from the haloarchaeon Haloarchaeobius sp. FL176.

Extracellular proteases of haloarchaea can adapt to high concentrations of NaCl and can find useful applications in industrial or biotechnology processes where hypersaline conditions are desired. The diversity of extracellular proteases produced by haloarchaea is largely unknown though the genomes of many species have been sequenced and are publicly available. In this study, a gene encoding the extracellular protease Hly176B from the haloarchaeon Haloarchaeobius sp. FL176 was cloned and expressed in Escherichia coli. A related gene homolog to hly176B, hly176A, from the same strain was also expressed in E.coli, but did not show any proteinase activity after the same renaturation process. Therefore, we focus on the enzymatic properties of the Hly176B. The catalytic triad Asp-His-Ser was confirmed via site-directed mutagenesis, indicating that Hly176B belongs to the class of serine proteases (halolysin). Unlike previously reported extracellular proteases from haloarchaea, the Hly176B remained active for a relatively long time in an almost salt-free solution. In addition, the Hly176B displayed prominent tolerance to some metal ions, surfactants and organic solvents, and exerts its highest enzyme activity at 40 °C, pH 8.0 and 0.5 M NaCl. Therefore, this study enriches our knowledge of extracellular proteases and expands their applications for various industrial uses.

Halocatena salina sp. nov., a filamentous halophilic archaeon isolated from Aiding Salt Lake.

A filamentous cell-shaped halophilic archaeon (strain AD-1T) was isolated from Aiding Salt Lake, PR China. Its colonies on HCM7 agar plates were pinkish white, 1-4 mm (diameter), elevated and round. The optimum conditions for growth were observed at 42 °C, 4.3 M NaCl, 0.01 M MgCl2 and pH 7. Strain AD-1T could hydrolyse Tween 60, Tween 80, starch and gelatin. Phylogenetic analysis based on 16S rRNA gene, rpoB' and the concatenated 484 single-copy orthologous proteins revealed that strain AD-1T formed a clade with Halocatena pleomorpha SPP-AMP-1T. The average nucleotide identity and in silico DNA-DNA hybridization values between strain AD-1T and Halocatena pleomorpha SPP-AMP-1T were both below the species delineation thresholds (95~96 and 70 %, respectively). The major phospholipids of strain AD-1T were phosphatidic acid, phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, while the major glycolipids were sulphated galactosyl mannosyl glucosyl diether, galactosyl mannosyl glucosyl diether and glucosyl mannosyl glucosyl diether. The phenotypic, phylogenetic and genome-based analyses suggested that strain AD-1T (=CGMCC 1.13724T=JCM 32960T) represents a novel species, for which the name Halocatena salina sp. nov. is proposed.

Halobaculum rubrum sp. nov., an extremely halophilic archaeon isolated from a salt lake.

A halophilic archaeal strain, designated C46T, was isolated from an inland salt lake in Qinghai Province, PR China. Results of phylogenetic analysis based on 16S rRNA gene sequences indicated that strain C46T belongs to the genus Halobaculum, and the closest phylogenetic relative is Halobaculum gomorrense DSM 9297T with 97.7 % similarity. Despite this, strain C46T was more related to Halobaculum saliterrae WSA2T than other members of the genus Halobaculum based on genome comparison and analysis, and the average nucleotide identity, in silico DNA-DNA hybridization, amino acid identity and percentage of conserved protein values between the two strains were 89.1, 53.3, 89.2 and 75.6 %, respectively, which are lower than the cutoff values proposed for species delimitation. The physiological, biochemical, genetic and genomic characteristics of strain C46T were different from those of its closest phylogenetic neighbours, which indicated that this strain represents a novel species of the genus Halobaculum, for which the name Halobaculum rubrum sp. nov. is proposed. The type strain is C46T (=CGMCC 1.13737T=JCM 32959T).

Induction of the antioxidant defense system using long-chain carotenoids extracted from extreme halophilic archaeon, Halovenus aranensis.

The field of microbial pigments is an emerging area in natural products science. Carotenoids form a major class of such pigments and are found to be diversely synthesized by microorganisms that reside in hypersaline ecosystems to provide resistance against oxidative stress. Human cells can benefit from compounds such as carotenoids as antioxidant agents through either their capability to quench free radicals or their effect on promoting the antioxidant defense pathway. In this study, the antioxidant effectiveness of carotenoid extract from an extremely halophilic archaeon Halovenus aranensis strain EB27T has been evaluated using different approaches. Finally, the ability of the extracted pigment to induce the antioxidant defense pathway of human primary skin fibroblast cells was studied. Hvn. aranensis carotenoid extract exhibited strong effectiveness such that at 2 µg/ml, the carotenoid extract fully neutralized the oxidative stress of hydrogen peroxide at its EC50 based on MTT assay. Results from real-time PCR of relevant genes, luciferase bioreporter of oxidative stress, and the western blot analysis further confirmed the antioxidant capability of the carotenoids. It was also shown the carotenoid extract had more antioxidant activity compared to ß-carotene the same concentration. Results suggest the carotenoid extract from this archaeon to have high potential for clinical and industrial applications.

Halalkalirubrum salinum gen. nov., sp. nov., a halophilic archaeon isolated from a saline lake.

A novel extremely halophilic archaeon, strain N1521T, was isolated from a saline lake in Tibet, China. Cells of the strain were pleomorphic and Gram-stain-negative. It produced red pigments. Growth was observed at 4-42 °C (optimum, 37 °C), pH 7.0-10.5 (optimum, 8.0-9.5), NaCl 11%-25% (optimum, 15%) and in the presence of 0-0.1 M MgCl2 (optimum, 0.05 M) in aerobic conditions. The minimum NaCl concentration that prevented cell lysis was 2% (w/v). The major polar lipids of strain N1521T were phosphatidylglycerol sulfate, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol and an unidentified glycolipid. The DNA G + C content was 58.37 mol%. According to 16S rRNA gene sequence comparisons, strain N1521T revealed the highest sequence similarity to Haloprofundus halophilus NK23T (91.38%) and Halogranum amylolyticum TNN58T (91.00%), and low sequence similarities (< 91%) with other genera in the order Haloferacales. Phylogenetic analysis based on the 16S rRNA gene and rpoB' gene sequence showed that strain N1521T was distinct from the members of the order Haloferacales. The digital DNA-DNA hybridization, average nucleotide identity and average amino acid identity values calculated from whole genome-sequence comparison between strain N1521T and the members of the order Haloferacales were in the ranges of 15.1-18.2%, 68.8-73.0%, and 58.4-63.9%, respectively. Phylogenetic tree reconstructions based on the whole-genome sequences revealed that strain N1521T was closer to the members of the family Halorubraceae. Based on the data obtained, strain N1521T is thus considered to represent a novel species of a new genus within the family Halorubraceae, for which the name Halalkalirubrum salinum gen. nov., sp. nov. is proposed. The type strain is N1521T (= CGMCC 1.16693 = JCM 33785).

Characterization of 3-isopropylmalate dehydrogenase from extremely halophilic archaeon Haloarcula japonica.

3-Isopropylmalate dehydrogenase (IPMDH) catalyzes oxidative decarboxylation of (2R, 3S)-3-isopropylmalate to 2-oxoisocaproate in leucine biosynthesis. In this study, recombinant IPMDH (HjIPMDH) from an extremely halophilic archaeon, Haloarcula japonica TR-1, was characterized. Activity of HjIPMDH increased as KCl concentration increased, and the maximum activity was observed at 3.0 m KCl. Analytical ultracentrifugation revealed that HjIPMDH formed a homotetramer at high KCl concentrations, and it dissociated to a monomer at low KCl concentrations. Additionally, HjIPMDH was thermally stabilized by higher KCl concentrations. This is the first report on haloarchaeal IPMDH.

Characterization of a GlgC homolog from extremely halophilic archaeon Haloarcula japonica.

Glycogen synthesis in bacteria is mainly organized by the products of glgB, glgC, and glgA genes comprising the widely known glg operon. On the genome of extremely halophilic archaeon Haloarcula japonica, there was a gene cluster analogous to the bacterial glg operon. In this study, we focused on a GlgC homolog of Ha. japonica, and its recombinant enzyme was prepared and characterized. The enzyme showed highest activity toward GTP and glucose-1-phosphate as substrates in the presence of 2.6 m KCl and predicted to be work as "GDP-glucose pyrophosphorylase" in Ha. japonica.

Halomicrococcus hydrotolerans gen. nov., sp. nov., an extremely halophilic archaeon isolated from a subterranean salt deposit.

A halophilic archaeon, strain H22T, was isolated from a subterranean salt deposit sampled at Yunnan salt mine, PR China. Colonies of strain H22T were light pink-pigmented. Cells were coccus, non-motile, Gram-stain-negative, and did not lyse in distilled water. The strain was aerobic and grew at 20-55 °C (optimum, 37 °C), in the presence of 10-30 % (w/v) NaCl (20 %) and at pH 6.5-9.0 (pH 7.0). Mg2+ was required for growth (optimum, 0.005 M). Major polar lipids were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and sulfated mannosyl-glucosyl-glycerol diether-1. Sequence similarity search based on the multiple 16S rRNA genes (rrnA, rrnB and rrnC) of strain H22T revealed that it was most closely related to species of the genera Haloarchaeobius, Haladaptatus, Halorussus and Halorubellus with relative low sequence similarities (91.9-93.7 %). The strain, however, shared highest rpoB' gene sequence identities with Halorussus rarus TBN4T (90.8 % rpoB' gene sequence similarity). Phylogenetic trees based on 16S rRNA and rpoB' gene sequences revealed a robust lineage of the strain H22T with members of related genera of the family Halobacteriaceae. The DNA G+C content of strain H22T was 62.9 mol%. Genome-based analysis of average nucleotide identity (ANI) and in silico DNA-DNA hybridization (DDH) between strains H22T and its closest relative were equal or lower than 77.7 and 22.4 %, respectively, which were far below the threshold for delineation of a new species. Based on ANI values, in silico DDH, and distinct morphological and physiological differences from the previously described taxa, we suggest that strain H22T represents a novel species of a new genus within the family Halobacteriaceae, for which the name Halomicrococcus hydrotolerans gen. nov., sp. nov. is proposed. The type strain is H22T (=CGMCC 1.16291T=NBRC 113231T).

Salinigranum halophilum sp. nov., isolated from marine solar salterns.

Three halophilic archaeal strains, YJ-53T, ZS-5 and DYF38, were isolated from marine solar salterns located in different provinces of China. The three strains formed a single cluster (99.7-99.8 and 97.9-99.2 % similarities, respectively) that was separate from the current two members of Salinigranum (96.7-98.0 and 89.8-92.9 % similarities, respectively) on the basis of 16S rRNA and rpoB' gene sequence comparisons and phylogenetic analysis. Diverse phenotypic characteristics differentiated strains YJ-53T, ZS-5 and DYF38 from Salinigranum rubrum GX10T and Salinigranum salinum YJ-50-S2T. The major polar lipids of isolated strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and two major glycolipids chromatographically identical to mannosyl glucosyl diether and sulfated mannosyl glucosyl diether, detected in the current members of Salinigranum. The OrthoANI and in silico DNA-DNA hybridization (DDH) values between the three strains were in the range of 97.7-98.4 % and 80.3-86.1 %, respectively, much higher than the threshold values proposed as species boundaries (average nucleotide identity 95-96 % and in silico DDH 70 %), revealing that the three strains represent one species. Results of comparative OrthoANI and in silico DDH analyses of the strains described in this study with validly described members of the genus Salinigranum supported that strains YJ-53T (=CGMCC 1.12860T=JCM 30238T), ZS-5 (=CGMCC 1.12867=JCM 30240) and DYF38 (=CGMCC 1.13779=JCM 33557) represent a novel species of the genus Salinigranum, for which the name Salinigranum halophilum sp. nov. is proposed.

Salinibaculum litoreum gen. nov., sp. nov., isolated from salted brown alga Laminaria.

A novel Gram-stain-negative, aerobic and rod-shaped halophilic archaeon, designated HD8-45T, was isolated from the red brine of salted brown alga Laminaria produced at Dalian, PR China. According to the results of 16S rRNA gene and rpoB' gene sequence comparisons, strain HD8-45T showed the highest sequence similarity to the corresponding genes of Salinirussus salinus YGH44T (95.1 and 85.2 % similarities, respectively), Halovenus aranensis EB27T (91.2 and 86.0 % similarities, respectively). The low sequence similarity and the phylogeny implied the novel generic status of strain HD8-45T. Genomic relatedness analyses showed that strain HD8-45T were clearly distinguished from other species in the order Halobacteriales, with average nucleotide identity, amino acid identity and in silico DNA-DNA hybridization values not more than 75.1, 65.6 and 21.5 %. The polar lipid pattern contained phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, two major glycolipids and two minor glycolipids. The two major glycolipids and a minor glycolipid were chromatographically identical to disulfated mannosyl glucosyl diether, sulfated mannosyl glucosyl diether and mannosyl glucosyl diether, respectively. The major respiratory quinones were menaquinone MK-8 and MK-8(H2). The DNA G+C content was 62.0 mol% (Tm) and 61.9 mol% (genome). All these results showed that strain HD8-45T represents a novel species of a new genus in the order Halobacteriales, for which the name Salinibaculum litoreum gen. nov., sp. nov. is proposed. The type strain of Salinibaculum litoreum is HD8-45T (=CGMCC 1.15328T=JCM 31107T).

Halorussus halophilus sp. nov., A Novel Halophilic Archaeon Isolated from a Marine Solar Saltern.

The halophilic archaeal strain ZS-3T (= CGMCC 1.12866T = JCM 30239T) was isolated from a sediment sample of Zhoushan marine solar saltern, P. R. China. Phylogenetic analyses based on 16S rRNA, rpoB' genes and the concatenation of 738 protein sequences reveal that strain ZS-3T was related to members of the genus Halorussus. The OrthoANI and in silico DDH values between strain ZS-3T and the current Halorussus members are much lower than the threshold values proposed as the species boundary (ANI 95-96% and in silico DDH 70%), suggesting that strain ZS-3T represents a novel species of Halorussus (Halorussus halophilus sp. nov.). Diverse phenotypic characteristics differentiate strain ZS-3T from current Halorussus members. Since the strain expressed diverse hydrolyzing enzyme activity, its complete genome was sequenced. The genome of strain ZS-3T was found to be 4,450,731 bp with total GC content of 61.51%, and comprises one chromosome and three plasmids. A total of 4694 protein coding genes, 43 tRNA genes and 6 rRNA genes were predicted. A CRISPR-Cas system was also detected. The genome encodes sixteen putative glycoside hydrolases, nine extracellular proteases, seventeen aminopeptidases, seven carboxypeptidases, one esterase and one nitrite reductase. The exploration of the hydrolase genes may expand our understanding of adapted mechanism of halophilic archaea surviving optimally in hypersaline environments where containing organic matter. Meanwhile, various hydrolyzing enzymes may extend this microorganism for further applications in salt-based fermentation.

Haloarcula sebkhae sp. nov., an extremely halophilic archaeon from Algerian hypersaline environment.

A halophilic organism, SWO25T, was isolated from water sampled in Algeria at the salt lake (sebkha) of Ouargla. The novel strain stained Gram-negative, and cells were pleomorphic with a red pigmentation. Strain SWO25T grew optimally at 35-45 °C, at pH 6.0-8.0 and 0.05-0.25 M MgCl2 concentrations. Cells were extremely halophilic, with optimal growth at 4.3-5.1 M NaCl. The predominant membrane polar lipids were C20C20 glycerol diether derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate, phosphatidylglycerol sulfate, triglycosyl diether and diglycosyl diether. The major respiratory menaquinone component was MK-8. Cells were highly tolerant to the presence of decane and isooctane in the growth medium. Chemotaxonomic properties supported the assignment of strain SWO25T to the genus Haloarcula. The DNA G+C content was 61.1mol%. DNA-DNA hybridization and phylogenetic analyses of the 16S rRNA and rpoB' genes showed that strain SWO25T is distinct from known Haloarcula species. Based on phenotypic, chemotaxonomic, genotypic and phylogenetic data, we describe a novel species of the genus Haloarcula, for which the name Haloarculasebkhae sp. nov. is proposed. The type strain is SWO25T (=CIP 110583T=JCM 19018T).

Halorussus litoreus sp. nov., isolated from the salted brown alga Laminaria.

A halophilic archaeal strain, designated HD8-51T, was isolated from the salted brown alga Laminaria. Cells of strain HD8-51T were motile, pleomorphic coccoid or ovoid, and formed red-pigmented colonies on agar plates. Strain HD8-51T grew optimally at 3.1 M NaCl, 0.03 M MgCl2, 30 °C and pH 7.0. Cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 0.85 M. Based on phylogenetic analyses of the 16S rRNA and rpoB' genes, strain HD8-51T was most closely related to members of the genus Halorussus (92.3-95.6 % and 89.2-91.7% similarities, respectively). The average nucleotide identity values and in silico DNA-DNA hybridization values between strain HD8-51T and Halorussus rarus TBN4T were 81.69 and 24.5 %, respectively. The major polar lipids of strain HD8-51T were phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS) and five glycolipids, sulfated galactosyl mannosyl glucosyl diether (S-TGD-1), galactosyl mannosyl glucosyl diether (TGD-1), sulfated mannosyl glucosyl diether (S-DGD-1), mannosyl glucosyl diether (DGD-1) and diglycosyl diether (DGD-2). The DNA G+C content was 65.9 mol%. Based on phenotypic, chemotaxonomic and phylogenetic properties, strain HD8-51T represents a novel species of the genus Halorussus, for which the name Halorussus litoreus sp. nov. is proposed. The type strain is HD8-51T (=CGMCC 1.15333T=JCM 31109T).

Halostella limicola sp. nov., isolated from saline soil sampled at the Tarim Basin.

A halophilic archaeaon, strain LT12T, was isolated from saline soil sampled at the Tarim Basin, PR China. The novel strain stained Gram-negative, cells were rod-shaped, and formed light red-pigmented colonies on agar plate. Strain LT12T grew optimally at 3.1 M NaCl, 0.05 M MgCl2, 37 °C and pH 7.5. The cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 1.4 M. Based on the results of phylogenetic analyses of the 16S rRNA and rpoB' genes, strain LT12T was most closely related to Halostella salina CBA1114T(94.4-95.9  and 93.6 % similarities, respectively). The average nucleotide identity and in silico DNA-DNA hybridization values between strain LT12T and H. salina CBA1114T were 81.0 and 24.3 %, respectively. The major polar lipids of strain LT12T were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and three unidentified glycolipids. The DNA G+C content was 67.2 mol % (genome). Based on the phenotypic, chemotaxonomic and phylogenetic properties, strain LT12T represents a novel species of the genus Halostella for which the name Halostellalimicola sp. nov. is proposed. The type strain is LT12T (=CGMCC 1.14941T=JCM 30667T).

Haloprofundus halophilus sp. nov., isolated from the saline soil of Tarim Basin.

A novel halophilic archaeon, designated NK23T, was isolated from an inland saline soil sampled from Xinjiang, China. The cells of strain NK23T were observed to be pleomorphic, to stain Gram-negative and form red-pigmented colonies on agar plates. The strain can grow at 25-50 °C (optimum 37 °C), at 0.9-4.8 M NaCl (optimum 2.1 M), at 0-1.0 M MgCl2 (optimum 0.05 M) and at pH 6.5-9.5 (optimum pH 7.0). The polar lipids were found to be phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, mannosyl glucosyl diether, sulfated mannosyl glucosyl diether, and three minor unidentified glycolipids, which were chromatographically identical to those detected in Haloprofundus (Hpf.) marisrubri CGMCC 1.14959T. On the basis of 16S rRNA gene and rpoB' gene sequence similarities and phylogenetic analysis, strain NK23T was found to be related to Hpf. marisrubri CGMCC 1.14959T (97.8% and 94.1% similarities, respectively). The average nucleotide identity values and in silico DNA-DNA hybridization values between strain NK23T and Hpf. marisrubri SB9T were 85.22% and 29.3%, respectively. The DNA G+C content of the novel strain was determined to be 65.29 mol%. Based on the phenotypic and chemotaxonomic data, together with phylogenetic relationships, strain NK23T (= CGMCC 1.14944T = JCM 30670T) is considered to represent a new species of the genus Haloprofundus, for which the name Haloprofundus halophilus sp. nov. is proposed.

Salinirubellus salinus gen. nov., sp. nov., isolated from a marine solar saltern.

A halophilic archaeal strain, designated ZS-35-S2T, was isolated from Zhoushan marine solar saltern in Zhejiang Province, China. Cells were pleomorphic, Gram-stain-negative and formed red-pigmented colonies on agar plates. The cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 8 % (w/v). Strain ZS-35-S2T was able to grow at 25-50 °C (optimum, 37 °C), with 1.4-4.8 M NaCl (optimum, 2.1 M), with 0-1.0 M MgCl2 (optimum, 0.1 M) and at pH 5.0-9.5 (optimum, pH 7.5). Phylogenetic tree reconstructions based on 16S rRNA genes and rpoB' genes revealed that strain ZS-35-S2T was distinct from the related genera Halomarina, Natronomonas, Halorientalis, Salinirubrum and Halobaculum of the order Halobacteriales. The major polar lipids of the strain were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and three unidentified glycolipids. The two major glycolipids were chromatographically identical to S-DGD-1 and DGD-1, respectively. The DNA G+C content of strain ZS-35-S2T is 67.0 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain ZS-35-S2T (=CGMCC 1.12551T=JCM 30036T) represents a novel species of a new genus within the order Halobacteriales, for which the name Salinirubellus salinus gen. nov., sp. nov. is proposed.

Halomarina rubra sp. nov., isolated from a marine solar saltern.

Halophilic archaeal strain ZS-47-ST was isolated from Zhoushan marine solar saltern, China. Cells were pleomorphic, stained Gram-negative, and formed red-pigmented colonies on agar plate. Strain ZS-47-ST was able to grow at 20-50 °C (optimum 37 °C), at 0.9-4.8 M NaCl (optimum 3.4 M), at 0.005-1.0 M MgCl2 (optimum 0.03 M), and at pH 5.5-9.5 (optimum pH 6.5-7.5). The cells lysed in distilled water and the minimal NaCl concentration to prevent cell-lysis was 5% (w/v). The major polar lipids were C20C20 and C20C25 diether derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, glucosyl mannosyl glucosyl diether, and three unidentified glycolipids. The 16S rRNA gene and rpoB' gene of strain ZS-47-ST were phylogenetically related to the corresponding genes of Halomarina oriensis JCM 16495T (98.57 and 92.94% similarities, respectively) and Halomarina salina CGMCC 1.12543T (97.96 and 93.65% similarities, respectively). The DNA G + C content of strain ZS-47-ST was 64.6 mol % (T m). The phenotypic, chemotaxonomic, and phylogenetic properties suggested that strain ZS-47-ST (=CGMCC 1.12563T = JCM 30037T) represents a new species of Halomarina, for which the name Halomarina rubra sp. nov. is proposed.

Haloplanus salinarum sp. nov., an extremely halophilic archaeon isolated from a solar saltern.

An extremely halophilic archaeal strain SP28T was isolated from the Gomso solar saltern, Republic of Korea. Cells of the new strain SP28T were pleomorphic and Gram stain negative, and produced red-pigmented colonies. These grew in medium with 2.5-4.5 M NaCl (optimum 3.1 M) and 0.05-0.5 M MgCl2 (optimum 0.1 M), at 25-50 °C (optimum 37 °C) and at a pH of 6.5-8.5 (optimum pH 8.0). Mg2+ was required for growth. A concentration of at least 2 M NaCl was required to prevent cell lysis. Polar lipids included phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and one glycolipid chromatographically identical to sulfated mannosyl glucosyl diether. 16S rRNA and rpoB' gene sequence analyses showed that strain SP28T is closely related to Haloplanus ruber R35T (97.3 and 94.1 %, 16S rRNA and rpoB' gene sequence similarity, respectively), Haloplanus litoreus GX21T (97.0 and 92.1 %), Haloplanus salinus YGH66T (96.0 and 91.9 %), Haloplanus vescus RO5-8T (95.9 and 90.9 %), Haloplanus aerogenes TBN37T (95.6 and 90.3 %) and Haloplanus natans RE-101T (95.3 and 89.8 %). The DNA G+C content of the novel strain SP28T was 66.2 mol%, which is slightly higher than that of Hpn.litoreus GX21T (65.8 mol%) and Hpn.ruber R35T (66.0 mol%). DNA-DNA hybridization values betweenHpn.ruber R35T and strain SP28T and between Hpn.litoreus GX21T and strain SP28T were about 24.8 and 20.7 %, respectively. We conclude that strain SP28T represents a novel species of the genus Haloplanus and propose the name Haloplanus salinarum sp. nov. The type strain is SP28T (=JCM 31424T=KCCM 43210T).

Halobacterium litoreum sp. nov., isolated from a marine solar saltern.

Halophilic archaeal strain ZS-54-S2T was isolated from Zhoushan marine solar saltern, China. Cells were rod-shaped, Gram-stain-negative and formed red-pigmented colonies on an agar plate. Strain ZS-54-S2T was able to grow at 20-50 °C (optimum 35 °C), at 1.7-4.8 M NaCl (optimum 3.9 M), at 0.005-1.0 M MgCl2 (optimum 0.05 M) and at pH 5.0-9.5 (optimum pH 7.0). The cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was found to be 5 % (w/v). The major polar lipids of the strain were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, two glycolipids, which were chromatographically identical to sulfated galactosyl mannosyl galactofuranosyl glucosyl diether and galactosyl mannosyl glucosyl diether, and an unidentified glycolipid, which was chromatographically identical to one detected in Halobacterium salinarum ATCC 33171T. The 16S rRNA gene and rpoB' gene of strain ZS-54-S2T were phylogenetically related to the corresponding genes of Halobacterium noricense JCM 15102T (97.5 % and 90.6 % relatedness, respectively), Halobacterium jilantaiense CGMCC 1.5337T (96.9 and 91.2 %), Halobacterium rubrum CGMCC 1.12575T (96.8 and 90.3 %) and Halobacterium salinarum CGMCC 1.1958T (96.5 and 88.4 %). The DNA G+C content of strain ZS-54-S2T was 66.7 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain ZS-54-S2T (=CGMCC 1.12562T=JCM 30038T) represents a new species of Halobacterium, for which the name Halobacteriumlitoreum sp. nov. is proposed.